Abstract This Special Instrumentation Grant proposal requests support for the purchase of a PerkinElmer IVIS Lumina S5 in vivo animal imaging system for shared use. The Lumina S5 enables researchers to anesthetize the animal and acquire optical 2D images from bioluminescent and fluorescent sources. Multiple animals (e.g. up to 5 mice) can be scanned simultaneously, enabling robust data acquisition on anatomical and molecular aspects of disease in vivo. The IVIS Lumina S5 also includes state of the art spectral unmixing features for sensitive multispectral imaging to monitor multiple biological events in the same animal. Access to this advanced in vivo imaging platform will enhance the research capabilities of our diverse, NIH-funded users group, which encompasses a broad range of projects with direct relevance to human health. These studies include rodent models of infectious disease (HIV/AIDS, influenza, Staphylococcus aureus, and Vibrio cholerae), host and pathogen gene expression, cancer research (glioblastoma and myeloid leukemia), and in vivo validation of new drug targets for obesity and fatty liver disease. The real-time imaging and analytical power of this instrument in live animals is not attainable by other means. The ability to monitor disease progression or treatment modalities in the same animal cohort as a function of time will also substantially reduce animal use and research costs. This instrument proposal has the full support of the University of Pittsburgh senior administration and will undoubtedly impart a substantial positive benefit on research progress now and well into the future.